JAMIA HAMDARD UNIVERSITY

PROJECT REPORT
Microorganism: Bacillus coagulans
(and it’s applications in food industry)

Submitted by: Submitted to:

Sarah Izhar. Dr. Shweta Joshi
Roll no: 25
Course: B.tech food technology
Year: 3rd (6th Sem)
INTRODUCTION

Bacillus coagulans is a good bacteria, termed as a probiotic. It has the ability to produce
lactic acid but is not similar to the probiotic Lactobacillus. Bacillus coagulans (B.coagulans)
was initially isolated from spoiled milk. In the year 1933, Horowitz-Wlassowa and
Nowotelnow identified it as lactobacillus sporogenes. Later it was named as B.coagulans.
Bacillus coagulans came into focus of the researchers and food manufacture, as it posses
features of both bacillus and Lactobacillus genera.

CHARACTERISTIC FEATURES

B.coagulans is known to be a gram positive bacteria, non-pathogenic and spore forming

bacteria, facultative anaerobes and lactic acid producing bacteria. It is a heat resistant
bacteria. The optimum growth temperature and optimum growth PH for B.coagulans is 35 to
60 degree Celsius and 5.5 to 6.5 respectively. B.coagulans is used as probiotics.
Although this microorganism produces acid, it does not emit gas from raffinose, mannitol,
maltose and sucrose fermentation. It has been known that B.coagulans causes spoilage in
dairy fruits and vegetable products due to acid production.
In addition to the production of lactic acid, thermostable alpha-amylase is also produced by
some strains. That is why B.coagulans is vital from an industrial point of view. The spores of
B.coagulans are terminal, while other species have spores which are central or sub terminal.
Furthermore, B.coagulans does not convert nitrate to nitrite and so it differs from other
bacillus species due to the absenteeism of Cytochrome-C oxidase. B.coagulans cannot grow
at PH 4.5 at a temperature of 65 degree Celsius and it was extracted from products that
contain milk and carbohydrates.

STATUS

It has been reported as safe by Food and Drug Administration (FDA) and European Union
Food Safety Authority (EFSA) and it comes under the list of Generally Recognised as Safe
(GRAS) and Qualified Presumption Safety. When B.coagulans GBI-30, 6086 was inspected
no risky or unsafe genes were found. Bacillus species including B.coagulans, B.subtilis var
natto, were identified as safe for human consumption.

GROWTH MEDIUM

The growth of bacillus coagulans is done using Molasses as a nutrient source. Molasses is a
carbon source which is available easily. It is a cheap nutrient source for the growth of
microbes. Distinct quality and concentration of molasses can influence the inoculum’s rate of
growth. MRS (de Man, Rogosa and Sharpe) broth can also be used for the growth of bacillus
coagulans.
This microorganism mainly generates spores in such conditions (culture conditions), that the
culture liquid constitutes some amount of inorganic salts and nitrogen containing organic
nutrient with very small amounts of fermentative saccharides. The liquid has a PH of 7.0 and
the culture is carried at a temperature of 37 to 50 degree celsius under anaerobic conditions
for about 2-4 days.

PROBIOTIC ACTIVITY OF bacillus coagulans

Food products that are heat-treated are not used for probiotic purposes because of the factors
affecting their stability and viability. To preclude this problem , B.coagulans due to their
heat-resistant spore forms could be used as probiotics. There are a few research studies on the
use of Bacillus species in human nutrition. There are a number of food products that contain
B.coagulans and these have been sold in different parts of the world. Probiotic
microorganism (ie;B.coagulans) have been used in probiotic food supplements, as freeze-
dried in dairy products such as yoghurt and also in fermented beverages. (Table 1 shows the
probiotic food supplements containing bacillus coagulans.)
The retention rates of Lactobacillus strains are influenced by the production process, storage
and transportation of food. Few strains of bacillus coagulans can survive in heat-treatments
done at a high temperature and can also survive in stomach condition than other commercial
probiotic micro organisms. Strain that posses such properties survive better in the

digestive tract.
 Table 1: Probiotic food supplements containing
bacillus coagulans

Source: https://www.researchgate.net/figure/Probiotic-food-supplements-containing-Bacillus-
coagulans_tbl1_325747096

APPLICATIONS OF bacillus coagulans IN FOOD INDUSTRY

In the past few years, the biological production of several metabolites ( like ethanol, lactic
acid, fumaric acid, xylonix acid and other products) has drawn attention in comparison to
chemical production with petroleum materials. Lactic acid is the most common metabolite.
A number of substances that are produced by bacillus coagulans is shown in Table 2.

Table 2: Substances produced by bacillus coagulans

Source:
https://www.researchgate.net/publication/325747096_Potential_Use_of_Bacillus_coagulans_
in_the_Food_Industry

➢ B.coagulans is an optimal microorganism in the production of lactic acid as it has the

ability to ferment glucose and xylose to lactic acid. This is done under anaerobic
conditions at a temperature below 50 degree Celsius. In various research studies, the
production of lactic acid from B.coagulans was done with the help of coffee extract,
 wheat straw, sugarcane pulp, sorghum water, corn cob, lignocellulosic hydrolysate
and corn flour as substrate.
➢ In food industry, beta-galactosidase is used for lactose hydrolysis. This is found in
nature and is extracted from animals, plants and microorganisms. It is also used in
food and pharmaceutical industries.
➢ Beta-galactosidaseis produced by yeasts (Kluyveromyces lactis and Kluyveromyces
marxianus),moulds (Aspergillus niger and Aspergillus oryzae) and bacteria
(B.circulans, E.coli, Lactobacillus bulgaricus, Lactobacillus thermophile).
B.coagulans is an exception. Another important enzyme used in food industry is
alpha-galactosidase and it has a number of applications commonly in sugar
production (also in biotechnology and medicine). This enzyme can enable the
digestion of legumes for example; soya bean by removing gala tissues like raffinose
and stachyose from their structures and thereby improving the gelation capacities.
There are many studies that have reported the production of alpha-galactosidase from
B.coagulans.

➢ Bacillus coagulans has proteolytic activity. It has been several years since the enzyme
amylase is used. These are important hydrolyse enzymes produced from plants and
microorganisms. Lipase are also produced from microorganisms. It is used in food
industry as well as pharmaceutical, textile and cosmetic industry. Some bacillus spp .
are known to produce lipase. In earlier studies lipase was known to produce from
bacillus coagulans..
➢ Lactic-acid producing bacteria is used in starter cultures for the fermentation in
yoghurt, kefir, sauerkraut, sausage, sourdough bread. Nigerian “Ugba” and Korean
“Kimchi”. These bacteria produce certain enzymes which help in food production.
Example; insoluble isomerase enzyme. It is also used in a majority of dietary
supplements—either ideally or along with vitamins, minerals, probiotic etc. Bacillus
coagulans can be used in the following products:
Baked goods, breakfast cereals and other grains, gelatins and puddings, cheese,
soft candy, soups, fats and oils, frozen dairy, milk products, confectionary and
frosting, hard candy, sugar substitutes, snack foods, non-alcoholic beverages and
instant coffee and tea.
 1. YOGURT

Preparation of yogurt is done with a mixture of major amount of milk powder and minor
amount of bacillus coagulans spores (also called Lactobacillus thermophillus) which have
certain feature of requiring sustenance at the time of formation of spores, competent of high
speed acid formation, taking less amount of time for transition into germ cells after budding
process and being thermobiotic so as to kill or inhibit the growth of saprophytes (undesirable)
including spore forming saprophytes during yogurt fermentation present in yogurt which
decontaminate or spoil the yogurt.

Procedure:
Yogurt is prepared, by adding boiling water to the mixture of milk powder and spores of
bacillus coagulans in a container to kill or hold back the growth of saprophytes. This mixture
is then allowed to ferment and stand for about 7-8 hours at a temperature of 50-55 degree
celsius with the lid on the container.
Following are the characteristics of bacillus spores:

1. The spores of bacillus coagulans have the ability to survive in boiling water (100
degrees) for more than 2 minutes.
2. Spores have the ability to produce germ cells in a period not more than 5 hours.
3. These germ cells have the tendency, such that fermentation of lactic acid occurs
quickly within a period of 4 hours at 50 degree celsius.
4. Germ cells have the feature that the milk coagulation occurs in not more than 5 hours,
after formation of germ cells.
5. Germ cells have the ability to generate lactic acid in final medium in a concentration
greater than 0.8%.
6. Germs cells can produce a PH in medium of fermentation process below 4.0.
 Fig.1: Probiotic Greek yogurt
Source: https://normansdairy.com/normans-dairy-introduces-the-worlds-first-greek-yogurt-
with-the-probiotic-ganedenbc30/

2. BAKED GOODS

Baked goods include breads, cakes, pies, tarts, pizza, corn chip, pastry, cookies, dehydrated
vegetables and fruits etc. In other words, a baked good includes anything that contains flour,
also any product that is heated is referred to as a baked good for example; fats and oils.
Bacillus coagulans is widely used in food industry in the preparation of baked goods.

Procedure:
The baked good contains flour which consists of a base mix and water, then the flour is
mixed to form a batter or a dough. To this isolated bacillus coagulans bacterium is applied
which is in the form of spores.
 Fig.2 : Probiotic baked products
Source: https://www.foodbusinessnews.net/articles/6674-baking-with-probiotics

3. NON DAIRY MILK

This product can be prepared from plant sources like grains, nuts, seeds or powdered seeds,
legumes etc. Following is the preparation of plant-based milk:

Procedure:
a) Grains, legumes or seeds are soaked in water for 12 hours at least. By doing this the
seeds are softened which enables easy blending and produces a finished product. This
process is excluded if soft plant sources are used.
b) These soaked grains are drained.
c) Water is then added to it. The ratio of plant source to water varies for eg; 1:2 to 1:4 on
the basis of preference of milk consistency. The mixture is then blended until smooth
consistency is obtained.
d) This mixture is further strained to remove finer particles left in the product.
e) Desired amount of bacillus coagulans spores are added and mixed with milk to add
the probiotic component into the product.
f) Finally, the plant-based milk is refrigerated until further use.
 Fig.3: Probiotic plant-based milk
Source: https://connect.naturalproductsexpo.com/product/non-dairy-yogurt-drink-0

CONCLUSION

Interest of consumer is increasing due to a diverse change in food and health and also in
their consumption habits. Production of probiotic foods and the benefits of probiotic
microorganisms on health has a great significance for the consumers, industry and
research centres. Heat-resistance of probiotic bacillus spp spores can come up with
advantages for probiotic foods that are heat-treated. Bacillus coagulans is gaining
attention due to its resistance to strong gastric acid and high temperature. Also, it is more
resistant to antibiotics than other lactic acid bacteria. Furthermore, the products used in
food industry could be produced from Bacillus coagulans.
References

1. Cash, H., Bush, M. A., Lefkowitz, A. R., & Keller, D. (2015). U.S. Patent
Application No. 14/694,481
2. Farmer, S., Lefkowitz, A. R., Bush, M. A., & Maske, D. T. (2020). U.S.
Patent Application No. 16/543,491.
3. Hata, K. (1980). U.S. Patent No. 4,210,672. Washington, DC: U.S. Patent
and Trademark Office.
4. Konuray, G., & Erginkaya, Z. (2018). Potential use of Bacillus coagulans
in the food industry. Foods, 7(6), 92.
5. Sanadi, N. A., Fan, Y. V., Leow, C. W., Wong, J. H., Koay, Y. S., Lee, C. T., ...
& Sarmidi, M. R. (2017). Growth of bacillus coagulans using molasses as a
nutrient source. Chemical Engineering Transactions, 56, 511-516.